Search PubMed⌕ Search

Biomedical subjects

A R Zuberi

Publications and source records attributed to A R Zuberi.

22 records · Page 2Linked to original sources

Transcriptional organization of a cloned chemotaxis locus of Bacillus subtilis.

A cloned chemotaxis operon has been characterized. Thirteen representative che mutations from different complementation groups were localized on the physical map by recombination experiments. The use of integration plasmids established that at least 10 of these complementation groups within this locus are cotranscribed. An additional three complementation groups may form part of the same transcript. The direction of transcription and the time of expression were determined from chromosomal che-lacZ gene fusions. The promoter was cloned and localized to a 3-kilobase fragment. Expression of beta-galactosidase from this promoter was observed primarily during the logarithmic phase of growth. Three-factor PBS1 cotransduction experiments were performed to order the che locus with respect to adjacent markers. The cheF141 mutation is 70 to 80% linked to pyrD1. This linkage is different from that reported previously (G. W. Ordal, D. O. Nettleton, and J. A. Hoch, J. Bacteriol. 154:1088-1097, 1983). The cheM127 mutation is 57% linked by transformation to spcB3. The gene order determined from all crosses is pyrD-cheF-cheM-spcB.

Bacillus subtilis↗

The nucleotide sequence and gene organization of the gerA spore germination operon of Bacillus subtilis 168.

The nucleotide sequence of the second and third genes in the Bacillus subtilis spore germination locus, gerA, has been determined and the amino acid (aa) sequence was derived. Two open reading frames (ORFs), corresponding to genes II and III, encode 364-aa residue and 373-aa residue polypeptides, respectively. The gene II product, Mr 41,257, would contain long stretches of hydrophobic aa residues and may be a membrane protein; the gene III product, Mr 42,363, is relatively hydrophilic but possesses an apparent signal peptide for transfer across, and perhaps localisation on, a membrane. The ORFs for genes I and II overlap by eleven codons and the termination codon of gene II overlaps the initiation codon of gene III. Insertional inactivation experiments using integrational plasmids have indicated that the gerA locus is a single transcriptional unit. The expression of the gerA genes has been studied using a lacZ transcriptional fusion; they constitute a developmentally regulated operon.

Amino Acid Sequence↗

Identification of three complementation units in the gerA spore germination locus of Bacillus subtilis.

The gerA locus, mutations in which affect the germination response of spores to L-alanine and related amino acids, is contained within a 6-kilobase region of DNA cloned in phage and plasmid vectors. Fragments from this region, subcloned in the shuttle vector pHV33, were introduced into Bacillus subtilis, and their ability to complement chromosomal gerA mutations in a recE4 background was examined. Although the plasmids were somewhat unstable, it was possible to score complementation within spore-containing colonies on nutrient agar by their ability to reduce 2,3,5-triphenyltetrazolium chloride in an overlay. These studies have assigned the 10 gerA mutations tested to three complementation groups. An analysis of Tn1000 insertions into the cloned DNA of two relatively stable plasmids that together encompass the entire gerA region has identified more precisely the location and extent of the complementation units; recombination studies and in vitro mutagenesis were used to further delineate the extents of two of the units. The evidence suggests that the three complementation units are adjacent and that they are probably capable of separate transcription.

Bacillus subtilis↗